Di-(2-ethylhexyl) phthalate (DEHP) is ubiquitous in the environment and has been proposed to lead to reproductive disruption. In this study, we systematically investigated the effects of different doses of DEHP exposure on female hypothalamic–pituitary–gonadal axis development. Female Sprague–Dawley rats were gavaged with vehicle (corn oil) or DEHP (5 or 500 mg kg–1 day–1) during postnatal Days (PNDs) 22–28 or PNDs 22–70. Results demonstrated that the low and high doses of DEHP exerted opposite effects on puberty onset, circulating luteinising hormone, serum oestradiol and progesterone levels, with the low dose (5 mg kg–1) promoting and the high dose (500 mg kg–1) inhibiting these parameters. Significant dose-related differences were also found in the D500 group with longer oestrous cycle duration, lower ovarian/bodyweight ratio, fewer corpus lutea and more abnormal ovarian stromal tissue in comparison with the oil or D5 groups. Molecular data showed that the hypothalamic Kiss1 mRNA expression in the anteroventral periventricular but not in the arcuate nucleus significantly decreased in the D500 rats and increased in the D5 rats relative to the rats in the oil group. These findings suggested that the kisspeptin system is a potential target for DEHP to disrupt reproductive development and function. Di-(2-ethylhexyl) phthalate (DEHP) is ubiquitous in the environment. This study found that low and high doses of DEHP had opposite effects on female puberty onset, hypothalamic kisspeptin expression and reproductive hormones. These results contribute to the comprehensive evaluation of DEHP toxicity on the human reproductive system. [ABSTRACT FROM AUTHOR]